Microfluidic in-line labeling method for continuous-flow protease inhibition analysis
First Claim
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1. A method of performing a protease assay in a microfluidic system, the method comprising:
- (i) flowing a protease through at least one microscale cavity;
(ii) contacting the protease with a protease substrate in the at least one microscale cavity or an additional microscale cavity, thereby reacting the protease substrate with the protease and producing one or more products;
(iii) labeling the one or more products by flowing a labeling reagent into the at least one microscale cavity or into the additional microscale cavity to react with the one or more products and produce one or more labeled products.
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Abstract
Enzyme assays are performed in microfluidic devices including, e.g., in-line labeling, separation, and detection of assay products. In-line labeling allows assays, e.g., protease assays, to be performed in a continuous flow microfluidic format. Also included are microfluidic devices and integrated systems for performing in-line labeling in continuous flow enzyme assays.
176 Citations
46 Claims
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1. A method of performing a protease assay in a microfluidic system, the method comprising:
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(i) flowing a protease through at least one microscale cavity;
(ii) contacting the protease with a protease substrate in the at least one microscale cavity or an additional microscale cavity, thereby reacting the protease substrate with the protease and producing one or more products;
(iii) labeling the one or more products by flowing a labeling reagent into the at least one microscale cavity or into the additional microscale cavity to react with the one or more products and produce one or more labeled products. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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24. A method of labeling one or more products in a microfluidic system, the method comprising:
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(i) flowing one or more unlabeled components through at least one microfluidic cavity;
(ii) reacting the one or more unlabeled components with one or more unlabeled reagents in the at least one microscale cavity or in an additional microscale cavity, resulting in one or more unlabeled products; and
,(iii) labeling the one or more unlabeled products by flowing a heterocyclic aromatic carbamate compound into contact with the one or more unlabeled products in the at least one microscale cavity or in the additional microscale cavity, thereby reacting the heterocyclic aromatic carbamate compound with the one or more unlabeled products and producing one or more labeled products.
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25. A microfluidic device for in-line labeling, the device comprising a body structure having a plurality of microscale channels disposed therein, the plurality of channels comprising:
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(i) a main channel;
in which main channel at least one protease and at least one protease substrate are reacted to produce one or more unlabeled products;
(ii) a labeling channel region fluidly coupled to the main channel;
in which labeling channel region the one or more unlabeled products are labeled to produce one or more labeled products;
(iii) a detection channel region fluidly coupled to the main channel, in which detection channel region the one or more labeled products are detected;
(iv) at least a first source fluidly coupled to the main channel, which first source comprises an amine-derivatizing reagent, which reagent reacts with the one or more unlabeled products to produce the one or more labeled products. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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40. A system for performing in-line labeling in a microfluidic device, the system comprising:
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(i) a microfluidic device comprising a plurality of microscale channels disposed therein, the plurality of microscale channels comprising;
(a) a main channel;
in which main channel at least one protease and at least one protease substrate are reacted to produce one or more unlabeled products;
(b) a labeling channel region fluidly coupled to the main channel;
in which labeling channel region the one or more unlabeled products are labeled to produce one or more labeled products;
(c) a detection channel region fluidly coupled to the main channel, in which detection channel region the one or more labeled products are detected;
(d) at least a first source fluidly coupled to the main channel, which first source comprises an amine-derivatizing reagent, which reagent reacts with the one or more unlabeled products to produce the one or more labeled products;
(ii) a fluid direction system fluidly coupled to the microfluidic device, which fluid direction system transports one or more of;
the at least one protease, the at least one protease substrate, the one or more unlabeled products, the one or more labeled products, and the amine-derivatizing reagent, through the plurality of microscale channels;
(iii) a control system operably linked to the fluid direction system, which control system instructs the fluid direction system to transport one or more of;
the at least one protease, the at least one protease substrate, the one or more unlabeled products, the one or more labeled products, and the amine-derivatizing reagent, through the plurality of microscale channels; and
,(iv) a detection system positioned proximal to the detection channel region for detecting the one or more labeled products. - View Dependent Claims (41, 42, 43, 44, 45, 46)
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Specification